An Improved Genetically Encoded Red Fluorescent Ca2+ Indicator for Detecting Optically Evoked Action Potentials 英文参考文献.docVIP

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An Improved Genetically Encoded Red Fluorescent Ca2+ Indicator for Detecting Optically Evoked Action Potentials 英文参考文献.doc

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AnImprovedGeneticallyEncodedRedFluorescentCa2IndicatorforDetectingOpticallyEvokedActionPotentials英文参考文献

AnImprovedGeneticallyEncodedRedFluorescentCa2+ IndicatorforDetectingOpticallyEvokedAction Potentials MasamichiOhkura1*.,TakuyaSasaki1.,ChiakiKobayashi2,YujiIkegaya2,JunichiNakai1* 1BrainScienceInstitute,SaitamaUniversity,Saitama,Japan,2LaboratoryofChemicalPharmacology,GraduateSchoolofPharmaceuticalSciences,UniversityofTokyo, Tokyo,Japan Abstract Genetically encoded Ca2+ indicators (GECIs) are powerful tools to image activities of defined cell populations. Here, we developed an improved red fluorescent GECI, termed R-CaMP1.07, by mutagenizing R-GECO1. In HeLa cell assays, R- CaMP1.07exhibiteda1.5–2-foldgreaterfluorescenceresponsecomparedtoR-GECO1.Inhippocampalpyramidalneurons, 2+ R-CaMP1.07detectedCa transientstriggeredbysingleactionpotentials(APs)withaprobabilityof95%andasignal-to- noiseratio.7ataframerateof50Hz.TheamplitudesofCa transientslinearlycorrelatedwiththenumberofAPs.The 2+ expressionofR-CaMP1.07didnotsignificantlyaltertheelectrophysiologicalpropertiesorsynapticactivitypatterns.Theco- expression of R-CaMP1.07 and channelrhodpsin-2 (ChR2), a photosensitive cation channel, in pyramidal neurons demonstratedthatR-CaMP1.07wasapplicableforthemonitoringofCa2+transientsinresponsetoopticallyevokedAPs, becausetheexcitationlightforR-CaMP1.07hardlyactivatedChR2.Thesetechnicaladvancementsprovideanovelstrategy formonitoringandmanipulatingneuronalactivitywithsinglecellresolution. Citation:OhkuraM,SasakiT,KobayashiC,IkegayaY,NakaiJ(2012)AnImprovedGeneticallyEncodedRedFluorescentCa2+IndicatorforDetectingOptically EvokedActionPotentials.PLoSONE7(7):e39933.doi:10.1371/journal.pone.0039933 Editor:StefanoL.Sensi,UniversityG.D’Annunzio,Italy ReceivedApril27,2012;AcceptedMay29,2012;PublishedJuly10,2012 Copyright: ? 2012 Ohkura et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:Thisworkw